Interpretive Summary: The confused flour beetle is a pest of stored products, and it infests a wide range of food products, from flour and cereals to spices. The insect reduces food quality and is responsible for large economic losses every year. Although a number of methods for detection of stored-product pests are common and widely used, they are time consuming and expensive. Therefore, establishing molecular methods of detection of stored-product pests could provide a useful alternative method. We have attempted to establish methods of detection of the confused flour beetle based on molecular biology techniques of standard and real-time PCR. The PCR method proved to be reliable and sensitive for detection, and the method is a possible supplemental detection method for quantitative assessment of infestation level.

Technical Abstract:
Tribolium confusum (confused flour beetle) is a stored-product pest which contaminates a wide range of food products, from flour and cereals to spices. The insect reduces food quality and is responsible for large economic losses every year. Although a number of methods for detection of stored-product pests are common and widely used, they are time consuming and expensive. Therefore, establishing molecular methods of detection of stored-product pests could provide a useful alternative method. We have undertaken attempts to establish methods of detection of T. confusum based on molecular biology techniques of standard and real-time PCR. Total DNA of T. confusum and Tribolium castaneum (red flour beetle), used as a negative control, was isolated from insects and used as a template in standard and real-time PCR reactions. Specific primers have been designed on the basis of sequences of ITS fragment of rDNA and I subunit of mitochondrial cytochrome oxidase (mtCOI) of T. confusum available in the GenBank database. Standard PCR reactions with primers specific to the ITS fragment proved to be reliable and sensitive. Real-time PCR reactions with primers specific for mtDNA are considered to serve as a supplemental detection method for quantitative assessment of the infestation level.